Authors thank Jeffrey Sellers to assist orthotopic pancreatic cancer mouse modeling. This work was supported by Research Initiative Pilot Awards from the Department of Radiology at UAB and NIH grants 2P30CA013148 and P50CA101955.

Tutte le procedure sono state approvate dalla cura e l&#39;uso degli animali Comitato Istituzionale presso la University of Alabama a Birmingham. Modellazione 1. ortotopico pancreatica Tumor mouse <ol><li> Cultura linee standard di cellule del pancreas-tumorali umane in mezzo modificato Eagle Dulbecco (DMEM) addizionato con siero fetale bovino 10%. Mantenere tutte le culture a 37 ° C in atmosfera umidificata con 5% di CO 2. </li><li> Utilizzare 8-10 femminili gravi topi immunodeficienti combinate settimana-vecchio. Posizionare gabbie per gli animali a 12 ore di luce e 12 ore ciclo buio a temperatura ambiente (21 ± 2 ° C), e il 60% di umidità. </li><li> Anestetizzare tutti gli animali con ventilazione con il 2% di isoflurano mescolato con l&#39;ossigeno (2 L / min) durante l&#39;intervento chirurgico. Confermare la profondità dell&#39;anestesia dal pizzico punta reflex. Posizionare animali su una piastra elettrica (37 ° C), per mantenere la temperatura corporea. Applicare veterinaria pomata oftalmica sugli occhi per prevenire la secchezza mentre sotto anestesia. &amp;# 160; </li><li> Rimuovere il capello nel quadrante superiore sinistro dell&#39;addome di ogni topo, e dare un farmaco analgesico (carpofen, 5 mg / kg di peso corporeo per via sottocutanea) nella zona. Applicare una soluzione betadine alla pelle esposta. Preparare strumenti chirurgici in autoclave. </li><li> Fare un 1 centimetro incisione nella pelle e del peritoneo con iris forbici diritte. Rimuovere delicatamente il pancreas dall&#39;addome con una pinzetta chirurgica. </li><li> Inserire 28 G ago di una siringa da 0,5 ml insulina nella coda del pancreas e poi lentamente infusione una soluzione di 2,5 milioni di cellule di cancro pancreatico umano in 30 ml di DMEM. Verificare che una piccola bozza viene creato nella testa del pancreas dalla soluzione. </li><li> Posizionare delicatamente il pancreas nuovamente dentro l&#39;addome con una pinzetta chirurgica. Chiudere il peritoneo e la pelle in 1 strato con 2 interrotto 5-0 suture Prolene, e quindi terminare la anestesia. Non restituire un animale che ha subito un intervento chirurgico per la compagnia di altri animali fino alla completa guarigione.60; Rimuovere suture a 7 ~ 10 giorni dopo l&#39;intervento chirurgico. </li><li> Dare un&#39;altra dose del farmaco analgesico (carpofen, 5 mg / kg di peso corporeo per via sottocutanea) a 24 ore dopo l&#39;intervento chirurgico. </li><li> Controllare la dimensione del tumore palpando l&#39;area chirurgia utilizzando due dita. I tumori in genere si sentono più denso e più irregolare rispetto ai tessuti circostanti e organi. Di solito ci vogliono ~ 1 - 2 settimane per iniziare a sentirsi un tumore. </li><li> Monitorare gli animali al giorno per i segni della malattia. Quando gli animali appaiono (normali comportamenti di governare e di evitamento mancanza) malati, abbiamo risolto utilizzando dislocazione cervicale mentre sotto anestesia. </li></ol> 2. Magnetic Resonance Imaging <ol><li> Applicare MRI quando le dimensioni del tumore è di circa 5-7 mm di diametro al solito 2 ~ 4 settimane dopo l&#39;impianto di cellule. Utilizzare uno scanner MR dedicato alle piccole immagini animale o uno scanner MR clinico dotato di una bobina specializzata per piccoli animali di imaging. NOTA: Abbiamo utilizzato un piccolo animale scanner RM 9.4T con una combinazione di un volume risuonatore 1 H/ Trasmettitore e un ricevitore bobina di superficie (30 mm di diametro) (Bruker BioSpin Corp., Billerica, MA). Una bobina di superficie fornisce migliore rapporto segnale-rumore (SNR) 14. </li><li> Preparare una risonanza magnetica agente di contrasto a base di gadolinio per iniettare ~ 0,1 - 0,2 mmol / kg per ogni animale in ~ 0,1 - 0,2 ml di PBS (tampone fosfato salino). NOTA: Abbiamo usato gadoteridolo e iniettato 0,2 mmol / kg in 0,15 ml di PBS in un periodo di 15 sec (0,1 ml / sec). </li><li> Preparare un tubo micro-polietilene (lunghezza: 7,62 mm, diametro interno: 0,28 millimetri, diametro esterno: 0,64 millimetri). Inserire un ago 30 G (lunghezza 12,7 mm) in una estremità del tubo, e una punta smussata dell&#39;ago 30 G (9,5 mm di lunghezza) nell&#39;altra estremità. Collegare una siringa da 1 ml contenente mezzo di contrasto MR all&#39;ago punta smussata, e spingere lentamente la siringa per riempire l&#39;intero tubo con l&#39;agente di contrasto MR. </li><li> Anestetizzare gli animali utilizzano ventilazione con ~ 1 - 2% di isoflurano mescolato con l&#39;ossigeno (2 L / min) per tutta la preparazione e l&#39;imaging. Confermare la profondità di Anesthesia by pizzico punta reflex. Applicare veterinaria pomata oftalmica sugli occhi per prevenire la secchezza mentre sotto anestesia. Dilatare la vena della coda con una lampada di calore prima dell&#39;inserimento dell&#39;ago. Afferra la metà dell&#39;ago 30 G con pinze Kelly, e inserirlo nella vena della coda. Tape sia la coda e il tubo su un foglio di plastica o di carta cartone (10 mm di larghezza x 100 mm di lunghezza) per mantenere la retta coda. </li><li> Posto l&#39;animale in posizione supina nel letto animale dotato di circolazione acqua calda (o aria calda) per regolare la temperatura corporea durante l&#39;imaging. Impostare la temperatura sul letto a 37 ° C. Inserire una sonda di temperatura rettale per controllare la temperatura corporea durante l&#39;imaging. </li><li> Applicare una scheda di plastica ortogonale piegata nella zona addominale. Assicurarsi che il tumore si trova dietro l&#39;estremità superiore della scheda, e poi tirare giù il bordo leggermente (~ 2 mm) per garantire il tumore e &#39;colto dal consiglio. Nastro il consiglio a letto animale saldamente. </li><li> Tape à transduce pad respirazioner (SA Instrument, Inc., Stony Brook, NY) sul petto per monitorare la respirazione degli animali durante l&#39;imaging. Inserire una bobina di superficie sulla sommità della regione tumorale, e nastro a letto animale saldamente. Premere il letto animale nello scanner MR per posizionare la regione tumorale al centro della bobina di volume (diametro interno 72 mm). </li><li> Eseguire adattamento e di accordo sia per il ricevitore e trasmettitore, seguito da spessori. </li><li> Inizia con una sequenza RM anatomica per individuare il tumore. Utilizzare una sequenza spin-echo (T2W) turbo T2-pesate per ottenere immagini assiali con i seguenti parametri di acquisizione. Tempo di ripetizione (TR) / tempo di echo (TE) = 3.000 / 34 msec, 128 x 128 a matrice, 30 x 30 mm campo visivo, il numero delle medie = 1, lunghezza del treno eco = 4, e 20 contigui 1 millimetro fette spesse in una modalità interlacciata per coprire l&#39;intera regione tumorale (tempo totale della scansione: 1,6 min). NOTA: Dal momento che i tumori pancreatici ortotopici sono più difficili da trovarsi di quelli sottocutanei, locali convenzionaliimmagini zer hanno risoluzione inferiore non possono essere utili. </li><li> Acquisire (T1W) immagini pesate in T1 con diversi angoli di vibrazione per recuperare T1 mappa. A questo scopo, utilizzare un gradiente echo approccio angolo multiflip con i seguenti parametri: tempo di ripetizione (TR) / tempo di eco (TE) = 115/3 msec, 128 x 128 matrice, un campo 30 x 30 mm di vista, il numero delle medie = 4, ~ 5-7 contigue 1 millimetro di spessore fette in una modalità interlacciata per coprire la regione tumorale, e sette angoli di vibrazione di 10, 20, 30, 40, 50, 60, e 70 (tempo totale della scansione per ogni angolo di vibrazione: 1 min). NOTA: Tuttavia, l&#39;angolo multiflip approccio è efficace solo quando il campo B1 omogeneità è alta. Se no, mappe T1 può essere ottenuto con l&#39;approccio TR multipla invece 15. </li><li> Acquisire le immagini T1W prima e dopo gadolinio basato iniezione di contrasto MR. Utilizzare gli stessi parametri di acquisizione e la geometria per la mappatura T1 ma con l&#39;angolo di vibrazione fisso di 30. Usa codifica lineare per garantire lo stato stazionario, quando si ottiene il centro di k-spazio, especially quando si utilizzano una breve TR e un angolo di vibrazione basso grado. Acquisire 5 immagini al basale prima iniezione di contrasto. Poi acquisire 40 immagini dopo l&#39;iniezione di contrasto (tempo totale della scansione: 45 min). Utilizzare una pompa a siringa per iniettare agente di contrasto ad un tasso costante (0,01 ml / sec). </li><li> Monitorare respirazione animale continuamente, e regolare la concentrazione isoflurano per mantenere la frequenza respiratoria di 50 - 100 respiri al minuto. Monitorare la temperatura corporea degli animali in tutta l&#39;imaging. </li><li> Dopo aver completato DCE-MRI, togliere l&#39;ago e altre sonde, e mettere l&#39;animale in una gabbia vuota letti con tovaglioli di carta. Massaggiare delicatamente la zona addominale inferiore. La gabbia deve essere posto metà sotto una lampada di calore per consentire all&#39;animale di muoversi dentro e fuori del gradiente termico come recupera. Non lasciare un animale incustodito fino a quando non ha ripreso conoscenza sufficiente per mantenere decubito sternale. </li></ol> 3. Image Processing and Analysis <ol><li> Regi tumorali Segmento<html> sulle immagini in T2w. In T2w immagini, l&#39;intensità del segnale in regione tumorale è luminoso di quello dei tessuti circostanti, così il confine tumore può essere delineata manualmente. NOTA: tecniche di segmentazione semi-automatiche come soglia globale o contornatura attivo possono essere utilizzati 16,17, ma l&#39;intensità sfondo uniforme devono essere corrette in particolare quando si utilizza una bobina di superficie. </li><li> Creare T1 e densità protonica mappe. Nelle immagini T1W acquisite con una sequenza gradient echo, supponendo che il tempo di eco (TE) è molto inferiore a T2 * valore, il valore del pixel è determinata dalla <img alt="Equazione 1" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq1.jpg" /> dove S 0 è la densità protonica, T 1 è costante T 1 tempo di rilassamento, TR è tempo di ripetizione, e θ è un angolo di vibrazione. L&#39;equazione (1) può essere riscritta per 641eq2.jpg &quot;/&gt; quando S (θ) / sinθ viene sostituito con Y, e S (θ) / tanθ viene sostituito con X. L&#39;equazione (2) è un&#39;equazione lineare, e il suo coefficiente angolare e può essere utilizzato per recuperare T1 e S 0 valori, rispettivamente. </li><li> Calcolare la concentrazione contrasto MR nelle immagini DCE-MR. Quando sulla base di contrasto MR gadolinio viene iniettato, T costante di tempo 1 relax è cambiata nel corso del tempo. Quindi, l&#39;equazione (1) può essere riscritto per <img alt="Equazione 3" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq3.jpg" /> T 1 (t) è correlata con la concentrazione contrasto MR, C (t), come segue, <img alt="Equazione 4" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq4.jpg" /> dove R 1 è relassività longitudinale del mezzo di contrasto MR. Quindi, combinando le equazioni (3) e (4), la concentrazione di contrasto MR è determinata <img alt = &quot;Equazione 5&quot; fo: content-width = src &quot;3in&quot; = &quot;/ files / ftp_upload / 52641 / 52641eq5.jpg&quot; /&gt; </li><li> Quantificare i parametri di farmacocinetica di MR contrasto agente. C p (t) presenta concentrazioni contrasto MR nel plasma sanguigno al tempo t dopo l&#39;inizio iniezione di contrasto. C p (t) è detta funzione di ingresso arteriosa (AIF). Se AIF è disponibile, i parametri farmacocinetici di mezzo di contrasto MR possono essere calcolati <img alt="Equazione 6" fo:content-width="4in" src="/files/ftp_upload/52641/52641eq6.jpg" /> dove C t (t) è la concentrazione contrasto MR in un tessuto bersaglio, v p è il volume del plasma sanguigno frazionale, v e è frazionata del volume extracellulare extravascolare, e K trans è costante trasferimento di volume. Costante di Flux, k ep, è pari a K trans diviso per v e. Se AIF non è disponibile, allora la regione modello di riferimento può essere utilizzato invece 18,19. La regione modello di riferimento si basa sul flusso limitato Kety modello 20 e usa la concentrazione contrasto in una regione di riferimento per eliminare la necessità di AIF come segue, <img alt="Equazione 7" fo:content-width="5in" src="/files/ftp_upload/52641/52641eq7.jpg" /> dove C t, ROI (t), K trans, ROI, ev e, ROI sono concentrazione contrasto, trasferimento volume costante, e il volume extravascolare-extracellulare frazionata, rispettivamente, nella regione di interesse (ROI), mentre C t, RR (t), K trans, RR, ev e, RR sono quelli della regione di riferimento. Muscoli paravertebrali è spesso selezionato come regione di riferimento, e v e, RR in pa murinomuscolo ravertebral si presume essere costante a 0,08 21. Abbiamo usato la regione modello di riferimento. </li></ol>

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	<li>Ergul, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+multifocality+and+axillary+nodal+involvement+in+early-stage+breast+cancer+patients+using+18F-FDG+PET/CT+compared+to+contrast-enhanced+and+diffusion-weighted+magnetic+resonance+imaging+and+sentinel+node+biopsy.">Assessment of multifocality and axillary nodal involvement in early-stage breast cancer patients using 18F-FDG PET/CT compared to contrast-enhanced and diffusion-weighted magnetic resonance imaging and sentinel node biopsy.</a> Acta Radiol. , (2014).</li><li>Park, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+early+response+to+concurrent+chemoradiotherapy+in+cervical+cancer:+value+of+diffusion-weighted+and+dynamic+contrast-enhanced+MR+imaging.">Assessment of early response to concurrent chemoradiotherapy in cervical cancer: value of diffusion-weighted and dynamic contrast-enhanced MR imaging.</a> Magn Reson Imaging. , (2014).</li><li>Nguyen, H. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prediction+of+chemotherapeutic+response+in+bladder+cancer+using+K-means+clustering+of+dynamic+contrast-enhanced+(DCE)-MRI+pharmacokinetic+parameters.">Prediction of chemotherapeutic response in bladder cancer using K-means clustering of dynamic contrast-enhanced (DCE)-MRI pharmacokinetic parameters.</a> J Magn Reson Imaging. 10, (2014).</li><li>Teo, Q. Q., Thng, C. H., Koh, T. S., Ng, Q. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+Contrast-enhanced+Magnetic+Resonance+Imaging:+Applications+in+Oncology.">Dynamic Contrast-enhanced Magnetic Resonance Imaging: Applications in Oncology.</a> Clin Oncol (R Coll Radiol). , (2014).</li><li>Zhang, X., Pagel, M. D., Baker, A. F., Gillies, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducibility+of+magnetic+resonance+perfusion+imaging.">Reproducibility of magnetic resonance perfusion imaging.</a> PLoS One. 9 (2), e89797 (2014).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pancreatic+adenocarcinoma:+a+pilot+study+of+quantitative+perfusion+and+diffusion-weighted+breath-hold+magnetic+resonance+imaging.">Pancreatic adenocarcinoma: a pilot study of quantitative perfusion and diffusion-weighted breath-hold magnetic resonance imaging.</a> Abdominal imaging. , (2014).</li><li>Derosier, L. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combination+Treatment+with+TRA-8+Anti+Death+Receptor+5+Antibody+and+CPT-11+Induces+Tumor+Regression+in+an+Orthotopic+Model+of+Pancreatic+Cancer.">Combination Treatment with TRA-8 Anti Death Receptor 5 Antibody and CPT-11 Induces Tumor Regression in an Orthotopic Model of Pancreatic Cancer.</a> Clin Cancer Res. 13 (18), 5535s-5543s (2007).</li><li>Derosier, L. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TRA-8+anti-DR5+monoclonal+antibody+and+gemcitabine+induce+apoptosis+and+inhibit+radiologically+validated+orthotopic+pancreatic+tumor+growth.">TRA-8 anti-DR5 monoclonal antibody and gemcitabine induce apoptosis and inhibit radiologically validated orthotopic pancreatic tumor growth.</a> Mol Cancer Ther. 6 (12), 3198-3207 (2007).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+therapy+evaluation+of+combined+anti-death+receptor+5+antibody+and+gemcitabine+in+orthotopic+pancreatic+tumor+xenografts+by+diffusion-weighted+magnetic+resonance+imaging.">Early therapy evaluation of combined anti-death receptor 5 antibody and gemcitabine in orthotopic pancreatic tumor xenografts by diffusion-weighted magnetic resonance imaging.</a> Cancer Res. 68 (20), 8369-8376 (2008).</li><li>Klein, S., Staring, M., Murphy, K., Viergever, M. A., Pluim, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=elastix:+a+toolbox+for+intensity-based+medical+image+registration.">elastix: a toolbox for intensity-based medical image registration.</a> IEEE Trans Med Imaging. 29 (1), 196-205 (2010).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+therapy+evaluation+of+combined+cetuximab+and+irinotecan+in+orthotopic+pancreatic+tumor+xenografts+by+dynamic+contrast-enhanced+magnetic+resonance+imaging.">Early therapy evaluation of combined cetuximab and irinotecan in orthotopic pancreatic tumor xenografts by dynamic contrast-enhanced magnetic resonance imaging.</a> Mol Imaging. 10 (3), 153-167 (2011).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antagonistic+effects+of+anti-EMMPRIN+antibody+when+combined+with+chemotherapy+against+hypovascular+pancreatic+cancers.">Antagonistic effects of anti-EMMPRIN antibody when combined with chemotherapy against hypovascular pancreatic cancers.</a> M Mol Imaging Biol. 16 (1), 85-94 (2014).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual+combination+therapy+targeting+DR5+and+EMMPRIN+in+pancreatic+adenocarcinoma.">Dual combination therapy targeting DR5 and EMMPRIN in pancreatic adenocarcinoma.</a> Mol Cancer Ther. 11 (2), 405-415 (2012).</li><li>Moyher, S. E., Vigneron, D. B., Nelson, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+coil+MR+imaging+of+the+human+brain+with+an+analytic+reception+profile+correction.">Surface coil MR imaging of the human brain with an analytic reception profile correction.</a> J Magn Reson Imaging. 5 (2), 139-144 (1995).</li><li>Voigt, T., Nehrke, K., Doessel, O., Katscher, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T1+corrected+B1+mapping+using+multi-TR+gradient+echo+sequences.">T1 corrected B1 mapping using multi-TR gradient echo sequences.</a> Magn Reson Med. 64 (3), 725-733 (2010).</li><li>Liu, H., Liu, Y., Zhao, Z., Zhang, L., Qiu, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+background+distribution-based+active+contour+model+for+three-dimensional+lesion+segmentation+in+breast+DCE-MRI.">A new background distribution-based active contour model for three-dimensional lesion segmentation in breast DCE-MRI.</a> Medical physics. 41 (8), 082303 (2014).</li><li>Sarkar, S., Das, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multilevel+image+thresholding+based+on+2D+histogram+and+maximum+Tsallis+entropy--a+differential+evolution+approach.">Multilevel image thresholding based on 2D histogram and maximum Tsallis entropy--a differential evolution approach.</a> IEEE Trans Image Process. 22 (12), 4788-4797 (2013).</li><li>Yankeelov, T. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+pharmacokinetic+analysis+of+DCE-MRI+data+without+an+arterial+input+function:+a+reference+region+model.">Quantitative pharmacokinetic analysis of DCE-MRI data without an arterial input function: a reference region model.</a> Magn Reson Imaging. 23 (4), 519-529 (2005).</li><li>Cardenas-Rodriguez, J., Howison, C. M., Pagel, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+linear+algorithm+of+the+reference+region+model+for+DCE-MRI+is+robust+and+relaxes+requirements+for+temporal+resolution.">A linear algorithm of the reference region model for DCE-MRI is robust and relaxes requirements for temporal resolution.</a> Magn Reson Imaging. 31 (4), 497-507 (2013).</li><li>Tofts, P. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimating+kinetic+parameters+from+dynamic+contrast-enhanced+T(1)-weighted+MRI+of+a+diffusable+tracer:+standardized+quantities+and+symbols.">Estimating kinetic parameters from dynamic contrast-enhanced T(1)-weighted MRI of a diffusable tracer: standardized quantities and symbols.</a> J Magn Reson Imaging. 10 (3), 223-232 (1999).</li><li>Yankeelov, T. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+a+reference+region+model+with+direct+measurement+of+an+AIF+in+the+analysis+of+DCE-MRI+data.">Comparison of a reference region model with direct measurement of an AIF in the analysis of DCE-MRI data.</a> Magn Reson Med. 57 (2), 353-361 (2007).</li><li>Cao, R. Y., Amand, T., Ford, M. D., Piomelli, U., Funk, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Murine+Angiotensin+II-Induced+Abdominal+Aortic+Aneurysm+Model:+Rupture+Risk+and+Inflammatory+Progression+Patterns.">The Murine Angiotensin II-Induced Abdominal Aortic Aneurysm Model: Rupture Risk and Inflammatory Progression Patterns.</a> Front Pharmacol. 1 (9), (2010).</li><li>Parker, G. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimentally-derived+functional+form+for+a+population-averaged+high-temporal-resolution+arterial+input+function+for+dynamic+contrast-enhanced+MRI.">Experimentally-derived functional form for a population-averaged high-temporal-resolution arterial input function for dynamic contrast-enhanced MRI.</a> Magn Reson Med. 56 (5), 993-1000 (2006).</li><li>Tseng, W., Leong, X., Engleman, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+mouse+model+of+colorectal+cancer.">Orthotopic mouse model of colorectal cancer.</a> J Vis Exp. (10), 484 (2007).</li><li>Bhullar, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+true+orthotopic+gastric+cancer+murine+model+using+electrocoagulation.">A true orthotopic gastric cancer murine model using electrocoagulation.</a> J Am Coll Surg. 217 (1), 64-70 (2013).</li></ol>

Authors do not have any conflicts of interest to declare.

Abbiamo introdotto metodi dettagliati di modellazione ortotopico tumore pancreatico utilizzando topi immunodeficienti, DCE-MRI dei tumori addominali nei topi, e la quantificazione dei suoi parametri cinetici. In ortotopico modellazione tumore pancreatico, bisogna fare attenzione quando si inserisce un ago nella coda del pancreas. In caso di successo, le cellule saranno trasferiti alla testa del pancreas creare una piccola bozza. Quando si applica una scheda di plastica ortogonalmente piegato, è fondamentale per conferm...

L&#39;obiettivo generale di questo metodo è quello di applicare dinamico di contrasto maggiore risonanza magnetica (DCE-MRI) per ortotopici xenotrapianti tumorali pancreatiche nei topi. DCE-MRI è un metodo non invasivo per valutare microcircolo in un tessuto bersaglio monitorando la variazione di MR contrasto per un certo periodo di tempo dopo l&#39;iniezione. DCE-MRI è stata utilizzata per diagnosticare tumori maligni e per valutare la risposta del tumore alle varie terapie 1-4. Quantitative DCE-MRI ha presentato alta riproducibilità 5. Per quantificare i parametri farmacocinetici di un agente di contrasto MR in un tessuto bersaglio, tutte le immagini DCE-MR acquisite in diversi momenti e T1 mappa ottenute prima iniezione di contrasto devono essere coregistrata 6. Tuttavia, a causa di movimenti respiratori e peristaltiche nella zona addominale, quantitativa DCE-MRI ha applicazione limitata per i tumori gastrointestinali. Ortotopico modelli tumorali pancreatiche sono stati utilizzati per valutareRisposta pancreatico-tumorale a seguito di terapie biologiche e chemioterapie 7,8. Modelli tumorali ortotopico sono considerate superiori ai modelli sottocutanei convenzionali, poiché il microambiente nel sito del tumore originale viene riflessa e risposta tumorale umano così alla terapia può essere previsto con maggiore precisione. Tuttavia, il pancreas del mouse si trova nel quadrante in alto a sinistra dell&#39;addome, in modo quantitativo DCE-MRI di ortotopici xenotrapianti tumorali pancreatiche nei topi non è stata immediatamente attuata. Abbiamo stabilito un protocollo di DCE-MRI dei tumori addominali nei topi fissando i tumori utilizzando una scheda di plastica ortogonalmente piegato per impedire il trasferimento di movimento della regione toracica 9. La pressione applicata da questa tavola è stato localizzato sulla zona addominale, e non ha portato a difficoltà respiratoria. Una tecnica automatizzata immagine coregistrazione è stato convalidato per DCE-MRI di organi addominali in una modalità senza respiro, ma esegue effectively solo quando le regioni obiettivo si muovono lentamente e regolarmente 10. La frequenza respiratoria di animali è variabile durante l&#39;imaging, ritenuta in modo fisico nella zona addominale sarà necessario recuperare i parametri farmacocinetici affidabili ortotopici modelli pancreatiche di topo tumore. Abbiamo quantificato con successo i parametri farmacocinetici di un agente di contrasto MR in ortotopici xenotrapianti tumorali pancreatiche usando la scheda di plastica ortogonalmente piegato in DCE-MRI 11-13. Ecco a voi la procedura dettagliata di ortotopica modellazione tumore pancreatico, DCE-MRI dei xenotrapianti tumorali nei topi, e la quantificazione dei parametri farmacocinetici.

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			<td height="21" style="height:21px;width:245px;">Name of Material/ Equipment</td>
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			<td style="width:119px;">Catalog Number</td>
			<td style="width:712px;">Comments/Description</td>
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			<td height="21" style="height:21px;">DMEM</td>
			<td>Invitrogen</td>
			<td>11965-118</td>
			<td></td>
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			<td height="21" style="height:21px;">Fetal bovine serum</td>
			<td>Harlan Laboratories</td>
			<td>BT-9501</td>
			<td></td>
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			<td height="21" style="height:21px;">Betadine</td>
			<td style="width:199px;">Purdue products</td>
			<td style="width:119px;">67618-153-01</td>
			<td></td>
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			<td height="21" style="height:21px;">5-0 Prolene sutures</td>
			<td style="width:199px;">Ethicon</td>
			<td style="width:119px;">8720H</td>
			<td></td>
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			<td height="42" style="height:42px;">9.4T MR scanner</td>
			<td>Bruker Biospin Corporation</td>
			<td style="width:119px;">BioSpec 94/20 USR</td>
			<td></td>
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			<td height="42" style="height:42px;">Gadoteridol</td>
			<td>Bracco Diagnostics Inc</td>
			<td style="width:119px;">NDC 0270-1111-03</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Micro-polyethelene tube</td>
			<td style="width:199px;">Strategic Applications, Inc</td>
			<td style="width:119px;">#PE-10-25</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">30G blunt tip needle</td>
			<td style="width:199px;">Strategic Applications, Inc</td>
			<td style="width:119px;">89134-194</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monitoring and gating system</td>
			<td style="width:199px;">SA instruments, Inc</td>
			<td style="width:119px;">Model 1030</td>
			<td>This is an MR compatiable system to measure resiratory rating and body temperature of small animals at the same time.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe pump</td>
			<td>New Era Pump Systems, Inc.</td>
			<td>NE-1600</td>
			<td></td>
		</tr>
	</tbody>
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dynamic contrast enhanced magnetic resonance imaging of an orthotopic pancreatic cancer mouse model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

Cellule tumorali pancreatiche umane crescere con successo nel topo pancreas creazione di un tumore solido. La figura 1 mostra le fotografie di (A) un normale pancreas dove soluzione delle cellule tumorali viene iniettato, e (B) un mouse rappresentante recante un ortotopico xenotrapianto tumore pancreatico (MIA PaCa-2 ). Tumor si trova nel quadrante superiore sinistro dell&#39;addome, accanto alla milza. Di solito ci vogliono 2-4 settimane per i tumori a crescere fino a 5-7 mm di diametro dopo l&#39;impi...

Watch this Scientific Journal Video about Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model at JoVE.com

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.

Contrasto dinamico migliorato risonanza magnetica di un modello ortotopico cancro del pancreas mouse

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe ...

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15.0K Views.  University of Alabama at Birmingham. The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy. 

Video: Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Contrast Enhanced Vessel Imaging using MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between different soft tissues and body fluids is inherently poor in micro-CT images 1. Under these circumstances, visualization of blood vessels becomes a nearly impossible task. To overcome this and to improve the visualization of blood vessels exogenous contrast agents can be used. Herein, we present a methodology for visualizing the vascular network in a rodent model. By using a long-acting aqueous colloidal polydisperse iodinated blood-pool contrast agent, eXIA 160XL, we optimized image acquisition parameters and volume-rendering techniques for finding blood vessels in live animals. Our findings suggest that, to achieve a superior contrast between bone and soft tissue from vessel, multiple-frames (at least 5-8/ frames per view), and 360-720 views (for a full 360&deg; rotation) acquisitions were mandatory. We have also demonstrated the use of a two-dimensional transfer function (where voxel color and opacity was assigned in proportion to CT value and gradient magnitude), in visualizing the anatomy and highlighting the structure of interest, the blood vessel network. This promising work lays a foundation for the qualitative and quantitative assessment of anti-angiogenesis preclinical studies using transgenic or xenograft tumor-bearing mice.

Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s).

Contrast Imaging nave migliorata utilizzando microCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between ...

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Medicina

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

Un modello ortotopico di cancro alla vescica Murine

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-fotone Imaging di invasione delle cellule tumorali in un modello di mouse ortotopico di carcinoma orale a cellule squamose

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

La radioterapia combinata in un modello di tumore ortotopico cervello di topo

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

Multi-modal Imaging di angiogenesi in un modello di ratto Nudo di metastasi ossee cancro al seno utilizzando la risonanza magnetica, tomografia computerizzata volumetrica e Ultrasuoni

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer cells into immunodeficient mice. Subcutaneous xenograft models have been valuable for preclinical screening and evaluation of new therapeutic treatments. There are a number of advantages to using a subcutaneous model; 1) rapid, 2) reproducible, and 3) tumor establishment, growth, and response to therapeutic agents may be monitored by visual inspection. However, substantial evidence has shed light on the short-comings of subcutaneous xenograft models1-3. For instance, medicinal treatments demonstrating curative properties in subcutaneous xenograft models often have no notable impact on the human disease. The microenvironment of the site of xenographic transplantation or injection lies at the heart of this dissimilarity.
Orthotopic tumor xenograft models provide a more biologically relevant context in which to study the disease. The advantages of implanting diseased cells or tissue into their anatomical origin equivalent within a host animal includes a suitable site for tumor-host interactions, development of disease-related metastases and the ability to examine site-specific influence on investigational therapeutic remedies. Therefore, orthotopic xenograft models harbor far more clinical value because they closely reproduce human disease. For these reasons, a number of groups have taken advantage of an orthotopic thyroid cancer model as a research tool4-7.
Here, we describe an approach that establishes an orthotopic model for the study of anaplastic thyroid carcinoma (ATC), which is highly invasive, resists treatment, and is virtually fatal in all diagnosed patients. Cultured ATC cells are prepared as a dissociated cellular suspension in a solution containing a basement membrane matrix. A small volume is slowly injected into the right thyroid gland. Overall appearance and health of the mice are monitored to ensure minimal post-operative complications and to gauge pathological penetrance of the cancer. Mice are sacrificed at 4 weeks, and tissue is collected for histological analysis. Animals may be taken at later time-points to examine more advance progression of the disease. Production of this orthotopic mouse model establishes a platform that accomplishes two objectives: 1) further our understanding of ATC pathology, and 2) screen current and future therapeutic agents for efficacy in combating ATC.

Generation of an orthotopic mouse model of anaplastic thyroid carcinoma is described here. This technique employs surgical placement of human anaplastic thyroid cancer cells into the thyroid of immunodeficient mice, thus creating a more clinically relevant setting to study disease progression as well as screen innovative therapeutic interventions.

Un modello di mouse ortotopico di anaplastico della tiroide Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

Un ortotopico cancro della vescica Modello di Gene Studi di consegna

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Bioluminescente modello ortotopico di cancro al pancreas Progressione

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

Un ortotopico Modello murino di Human Prostate Cancer Metastasi

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

Simultanea Imaging PET / MRI Durante mouse cerebrale ipossia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...